Peripheral blood biomarkers for evaluating the antitumor immune effects of radiation therapy

Abstract

This invention provides a peripheral blood biomarker for evaluating the antitumor immune response to radiation therapy. This invention also provides a method for using the composition of cell subsets in a sample obtained from a subject as an indicator of immune activation following radiation therapy in the subject. The presence and / or extent of immune activation following radiation therapy in the subject can be determined by comparing the amount of CD4+ T cell subsets associated with dendritic cell stimulation under antitumor immune response, or dendritic cell subsets associated with dendritic cell stimulation under antitumor immune response, to a baseline.

Description

[Technical Field] This invention relates to the field of cancer treatment. In particular, it concerns the evaluation of the immune effects of radiation therapy in cancer treatment. [Previous Technology] Reports have described the regression of cancer cells outside the irradiated field of view during radiation therapy, also known as the abscopal effect. The abscopal effect is generally believed to be a phenomenon resulting from any immune activation; however, many aspects of the detailed mechanism remain unclear, and no biomarkers have been found to quantify or evaluate the abscopal effect. [Summary of the Invention] [Technical means to solve the problem] This invention provides a method for using the composition of cell subpopulations in a sample obtained from a subject as an indicator of immune activation induced by radiation therapy in the subject. The presence and / or degree of immune activation induced by radiation therapy in the subject can be determined by comparing the amount of specific cell subpopulations described in this specification with a benchmark. The cell subpopulations that can be used as indicators in this invention are not limited, but may include: CD4+ T cell subpopulations related to dendritic cell stimulation under antitumor immune responses, dendritic cell subpopulations related to dendritic cell stimulation under antitumor immune responses, or CD8+ T cell subpopulations related to dendritic cell stimulation under antitumor immune responses. For example, CD4+ T cell subpopulations related to dendritic cell stimulation under antitumor immune responses are cell subpopulations included in the CD62LlowCD4+ T cell population (e.g., the CD62LlowCD4+ T cell subpopulation itself, or the ICOS+CD62LlowCD4+ T cell subpopulation, etc.). For example, dendritic cell subpopulations related to dendritic cell stimulation under antitumor immune responses are HLA-DR+CD141+CD11c+ cell subpopulations, etc. For example, CD8+ T cell subpopulations related to dendritic cell stimulation under antitumor immune responses are CD137+CD62LlowCD8+ T cell subpopulations, etc. In another embodiment of the invention, the radiation-induced immune activation in the subject indicates whether or when cancer immunotherapy should be administered. It is generally believed that administering cancer immunotherapy utilizing the immune response to cancer is more advantageous at the point when radiation-induced immune activation occurs; however, to date, there is no biomarker for determining radiation-induced immune activation. As a cancer immunotherapy, examples that are preferable include cancer immunotherapy involving the administration of immune checkpoint inhibitors. The administration of immune checkpoint inhibitors can be combined with other forms of treatment, and can also be combined with different agents (e.g., different immune checkpoint inhibitors). Examples of embodiments of the present invention are shown in the following items. (Item 1) A method that uses the composition of cell subpopulations in a sample obtained from a subject who has received radiation therapy as an indicator of immune activation in the subject due to radiation therapy, and includes the step of analyzing the composition of cell subpopulations in the sample obtained from the subject, indicating the presence or absence of immune activation in the subject by comparing the amount of CD4+ T cell subpopulations in the sample that are associated with dendritic cell stimulation under antitumor immune response with a baseline. (Item 2) A method that uses the composition of cell subpopulations in a sample obtained from a subject who has received radiation therapy as an indicator of immune activation in the subject due to radiation therapy, and includes: analyzing the composition of cell subpopulations in the sample obtained from the subject, and indicating the presence or absence of immune activation in the subject by comparing the amount of dendritic cell subpopulations in the sample that are associated with dendritic cell stimulation under antitumor immune response to a baseline. (Item 3) A method that uses the composition of cell subpopulations in a sample obtained from a subject who has received radiation therapy as an indicator of immune activation in the subject due to radiation therapy, and includes: analyzing the composition of cell subpopulations in the sample obtained from the subject, and indicating the presence or absence of immune activation in the subject by comparing the amount of CD8+ T cell subpopulations in the sample that are associated with dendritic cell stimulation under antitumor immune response to a baseline. (Item 4) The method of any of the above items, wherein the presence or absence of immune activation in the subject is indicated by comparing at least two quantities of a subset of the above sample, consisting of the amount of a subset of CD4+ T cells selected for dendritic cell stimulation under antitumor immune response, the amount of a subset of dendritic cells selected for dendritic cell stimulation under antitumor immune response, and the amount of a subset of CD8+ T cells selected for dendritic cell stimulation under antitumor immune response, with a baseline. (Item 5) The method of any of the above items, wherein the CD4+ T cell subset of the above sample that is related to dendritic cell stimulation under antitumor immune response is a subset of the CD62LlowCD4+ T cell population. (Item 6) The method of any of the above items, wherein the CD4+ T cell subset of the above sample that is related to dendritic cell stimulation under antitumor immune response is a CD62LlowCD4+ T cell subset or a CCR7+CD45RA-CD62LlowCD4+ T cell subset. (Item 7) The method of any of the above items, wherein the CD4+ T cell subsets associated with dendritic cell stimulation under antitumor immune response are ICOS+CD62LlowCD4+ T cell subsets, LAG3+CD62LlowCD4+ T cell subsets, PD-1+CD62LlowCD4+ T cell subsets or CD28+CD62LlowCD4+ T cell subsets.(Item 8) The method of any of the above items, wherein the dendritic cell subset associated with dendritic cell stimulation under antitumor immune response is the HLA-DR+CD141+CD11c+ cell subset. (Item 9) The method of any of the above items, wherein the CD8+ T cell subset associated with dendritic cell stimulation under antitumor immune response is a cell subset contained in the CD62LlowCD8+ T cell population. (Item 10) The method of any of the above items, wherein the CD8+ T cell subset associated with dendritic cell stimulation under antitumor immune response is the CD137+CD62LlowCD8+ T cell subset. (Item 11) The method of any of the above items, wherein the sample is a peripheral blood sample. (Item 12) The method of any of the above items, wherein the reference is the amount of the cell subset in the sample of the subject prior to the above radiation therapy. (Item 13) As in any of the above items, an increase in the amount of the aforementioned cell subpopulation in the sample compared to the aforementioned baseline indicates the generation of immune activation by radiation therapy in the subject. (Item 14) As in any of the above items, no increase in the amount of the aforementioned cell subpopulation in the sample compared to the aforementioned baseline indicates the absence of immune activation by radiation therapy in the subject. (Item 15) As in any of the above items, the radiation therapy is performed within the irradiation area including the tumor. (Item 16) As in any of the above items, the generation of immune activation by radiation therapy in the subject indicates that cancer immunotherapy including an immune checkpoint inhibitor should be administered to the subject at that time point. (Item 17) As in any of the above items, the absence of immune activation by radiation therapy in the subject indicates that radiation therapy should be administered to the subject again. (Item 18) A method, as described in any of the preceding items, further comprising using the composition of cell subpopulations in samples obtained from a subject at a plurality of time points as an indicator for monitoring immune activation in the subject due to radiation therapy, and comprising the step of analyzing the composition of cell subpopulations in samples obtained from the subject at a plurality of time points. (Item 19) A composition comprising an immune checkpoint inhibitor for treating cancer in a subject, wherein the composition is administered to a subject who has received radiation therapy and for whom immune activation has been indicated by any of the preceding items. (Item 20) A composition of any of the preceding items, wherein the immune checkpoint inhibitor is a PD-1 inhibitor and / or a PD-L1 inhibitor. (Item 21) A composition of any of the preceding items, administered in combination with one or more other agents.(Item 22) A composition of any of the above items, wherein the additional agent comprises a second immune checkpoint inhibitor. (Item 23) An article comprising a composition of any of the above items and accompanying documentation describing the use of the composition in conjunction with radiation therapy. (Item 24) A kit for determining whether radiation-induced immune activation has occurred in a subject, comprising: a detection agent for (A) CD4 and CD62L; (B)(i) a marker selected from ICOS, PD-1, LAG-3 and CD28, (ii) CD4, and (iii) CD62L; (C) CD11c, CD141 and HLA-DR; (D) CD11c, CD123 and HLA-DR; or (E) CD8, CD62L and CD137. (Item 25) A composition characterized by comprising an immune checkpoint inhibitor for treating cancer, and administered to a subject who has received radiation therapy, the subject having an immune cell composition unresponsive to cancer immunotherapy prior to receiving radiation therapy. (Item 26) The composition of any of the above items, wherein the immune cell composition unresponsive to the cancer immunotherapy is the amount or ratio of a CD4+CD62Llow cell subset below a threshold. (Item 27) The composition of any of the above items, wherein the immune checkpoint inhibitor comprises an anti-PD-1 antibody and / or an anti-PD-L1 antibody. [Effects of the Invention]. This invention allows for the quantification or evaluation of immune activation induced by radiation therapy. This enables the long-term monitoring of priming phase activation and the increase in effector T cells resulting from radiation therapy. Furthermore, this invention allows for the determination of the appropriate timing for immune checkpoint (e.g., PD-1) inhibitory therapy following radiation therapy. Additionally, the biomarkers of this invention can serve as biomarkers for evaluating the effectiveness of radiation-induced vaccine therapy (which demonstrates T-cell priming effects) in converting patients previously ineffective to effective immune checkpoint inhibitors, and as indicators for selecting patients requiring treatment for such changes.

Implementation Method

Claims

1. A method that uses the composition of cell subsets in a sample obtained from a subject who has received radiation therapy as an indicator of radiation-induced immune activation in the subject, comprising: analyzing the composition of cell subsets in the sample obtained from the subject; indicating the presence or absence of immune activation in the subject by comparing the amount of a CD4+ T cell subset associated with dendritic cell stimulation under antitumor immune response in the sample with a baseline; wherein the CD4+ T cell subset associated with dendritic cell stimulation under antitumor immune response is a cell subset comprising the CD62L lowCD4+ T cell population; an increase in the amount of the cell subset in the sample compared to the baseline indicates radiation-induced immune activation in the subject; and no increase in the amount of the cell subset in the sample compared to the baseline indicates the absence of radiation-induced immune activation in the subject. Based on the immune activation generated in the subjects by radiation therapy, it is indicated that at that point in time, the subjects should be given cancer immunotherapy containing immune checkpoint inhibitors, and the immune checkpoint inhibitors are PD-1 inhibitors, PD-L1 inhibitors, and / or CTLA-4 inhibitors.

2. The method of claim 1, wherein the absence of immune activation by radiation therapy in the subject indicates that the subject should be subjected to radiation therapy again.

3. The method of claim 2, further comprising the step of analyzing the composition of the cell subpopulations in the sample obtained from the subject who has received re-radiation therapy.

4. The method of claim 1, wherein the presence or absence of immune activation in the subject is indicated by comparing a measure of a subset of the above-mentioned sample, consisting of the amount of a dendritic cell subset selected for dendritic cell stimulation in response to antitumor immune response and the amount of a CD8+ T cell subset selected for dendritic cell stimulation in response to antitumor immune response, with a baseline.

5. The method of any one of claims 1 to 4, wherein the CD4+ T cell subsets associated with dendritic cell stimulation under antitumor immune response are CCR7+CD45RA-CD62L lowCD4+ T cell subsets, ICOS+CD62L lowCD4+ T cell subsets, LAG3+CD62L lowCD4+ T cell subsets, PD-1+CD62L lowCD4+ T cell subsets, or CD28+CD62L lowCD4+ T cell subsets.

6. The method of claim 4, wherein the dendritic cell subset associated with dendritic cell stimulation under antitumor immune response is an HLA-DR+CD141+CD11c+ cell subset.

7. The method of claim 4, wherein the aforementioned CD8+ T cell subset associated with dendritic cell stimulation under antitumor immune response is a subset of the CD62L lowCD8+ T cell population.

8. The method of claim 7, wherein the CD8+ T cell subset associated with dendritic cell stimulation under antitumor immune response is the CD137+CD62L lowCD8+ T cell subset.

9. The method of claim 1, wherein the above-mentioned sample is a peripheral blood sample.

10. The method of claim 1, wherein the reference is the amount of the cell subpopulation in the sample of the subject prior to the radiation therapy.

11. The method of claim 1, wherein the above-mentioned radiation therapy is performed within the irradiation area containing the tumor.

12. The method of claim 1, wherein the immune checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, or a combination of an anti-PD-1 antibody or an anti-PD-L1 antibody and a CTLA-4 inhibitor.

13. The method of claim 1, wherein the composition of cell subpopulations in samples obtained from the subject at a plurality of time points is further used as an indicator for monitoring immune activation in the subject by radiation therapy, and includes the step of analyzing the composition of cell subpopulations in samples obtained from the subject at a plurality of time points.

14. Use of a composition comprising an immune checkpoint inhibitor for manufacturing a medicine for treating cancer in a subject, wherein the composition is administered to a subject who has received radiation therapy and whose immune activation is indicated by the method of claim 1.

15. As claimed in claim 14, wherein the immune checkpoint inhibitor is a PD-1 inhibitor, a PD-L1 inhibitor, or a CTLA-4 inhibitor.

16. As claimed in claims 14 or 15, wherein the above composition is administered in combination with one or more other pharmaceutical agents.

17. As claimed in claim 16, wherein the aforementioned additional agent contains a second immune checkpoint inhibitor.

18. An application of a kit for the method of claim 1, wherein the kit comprises the composition used in any of claims 14 to 17, and accompanying documentation describing the use of the composition in conjunction with radiation therapy.

19. An application of a kit to the method of claim 1, wherein the kit comprises: determining whether radiation-induced immune activation occurs in a subject for (A) CD4 and CD62L detection agents by comparing the amount of a CD4+ T cell subset associated with dendritic stimulation under antitumor immune response in a sample obtained from the subject with a baseline.

20. For the purposes of any of claims 14 to 17, wherein the subject, prior to receiving radiation therapy, has an immune cell composition that is unresponsive to cancer immunotherapy, wherein the unresponsive immune cell composition is below the amount or ratio of the CD62L lowCD4+ cell subset used to determine whether the subject is unresponsive to cancer immunotherapy.

21. As claimed in claim 17, wherein the immune checkpoint inhibitor comprises an anti-PD-1 antibody, an anti-PD-L1 antibody, and / or a CTLA-4 inhibitor.

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